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目的:探讨丙烯酰胺(AA)对人SH-SY5Y神经细胞凋亡和miR-21表达的影响。方法:不同浓度丙烯酰胺作用于人神经母细胞瘤SH-SY5Y细胞24 h后,采用四甲基偶氮唑盐(MTT)法测定丙烯酰胺对SH-SY5Y细胞活力的影响,Hoechst 33258荧光染色法检测细胞凋亡,real-time PCR检测miR-21表达水平,Western blot检测PTEN、p-AKT、AKT、Bcl-2、Bax、caspase 9和caspase 3蛋白表达。结果:丙烯酰胺剂量依赖性地降低SH-SY5Y细胞活性,Hoechst 33258染色显示明显细胞凋亡形态变化;丙烯酰胺显著降低SH-SY5Y细胞miR-21的表达,同时升高PTEN、Bax、caspase 9、caspase 3水平并降低p-AKT和Bcl-2表达。结论:丙烯酰胺可通过线粒体途径诱导SH-SY5Y神经细胞凋亡,其机制可能与丙烯酰胺引起miR-21表达下调有关。
Objective: To investigate the effect of acrylamide (AA) on the apoptosis of human SH-SY5Y cells and the expression of miR-21. Methods: The effect of acrylamide on the activity of SH-SY5Y cells was detected by MTT assay after 24 h treatment with different concentrations of acrylamide on human neuroblastoma SH-SY5Y cells. Hoechst 33258 fluorescence staining The expression of miR-21 was detected by real-time PCR and the protein expressions of PTEN, p-AKT, AKT, Bcl-2, Bax, caspase 9 and caspase 3 were detected by Western blot. Results: Acrylamide dose-dependently reduced the activity of SH-SY5Y cells, Hoechst 33258 staining showed obvious morphological changes of apoptotic cells. Acrylamide significantly decreased the expression of miR-21 and increased the expressions of PTEN, Bax and caspase 9, caspase 3 levels and reduce p-AKT and Bcl-2 expression. CONCLUSION: Acrylamide can induce apoptosis of SH-SY5Y neurons through mitochondrial pathway, which may be related to the down-regulation of miR-21 induced by acrylamide.